| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 17366 | Enzyme and Microbial Technology | 2013 | 7 Pages |
A sterol glucosyltransferase-encoded gene was isolated from Salinispora tropica CNB-440, a marine, sediment-dwelling, Gram positive bacterium that produces the potent anticancer compound, salinosporamide A. The full-length gene consists of 1284 nucleotides and encodes 427 amino acids with a calculated mass of 45.65 kDa. The gene was then cloned and heterologously expressed in Escherichia coli BL21(DE3). The amino acid sequence shares 39% similarity with the glycosyltransferase from Withania somnifera, which belongs to glycosyltransferase family 1. Enzyme reactions were carried out with the various free sterols (acceptor) and NDP-sugars (donor). The purified protein only showed activity for glucosylation of β-sitosterol with UDP-D-glucose and TDP-D-glucose donors, and optimal activity at pH 7.5 and 37 °C. Among these two donors, UDP-D-glucose was preferred.
Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► A sterol glucosyltransferase gene was isolated from Salinispora tropica CNB-440. ► Phylogenetic relationship analysis based on the C-terminal amino acid sequences. ► Heterologous expression successfully in E. coli BL21(DE3) and purified the protein. ► Biochemical properties were characterized and compared with another published reports.
